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Reproductive Biotechnology in Finfish Aquaculture

Reproductive Biotechnology in Finfish Aquaculture

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Reproductive Biotechnology in Finfish Aquaculture

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Dec 2, 2012


The successful reproduction of cultured brood stock is essential to the sustainable aquaculture of aquatic organisms. This book describes recent advances in the field of finfish reproductive biotechnology. The chapters in this volume are written by eminent scientists who review the progress and assess the status of biotechnology research that is applicable to the reproduction of finfish species for aquaculture. A wide range of topics is included starting with broodstock technologies such as broodstock genetics, broodstock nutrition, environmental control of maturation and impacts of stress on broodstock, gametes and progeny.

The volume includes technologies for induction of ovulation and spermiation using synthetic hypothalamic peptides. Gamete technologies which are described include cryopreservation, chromosome set manipulation, disease prevention and control for gametes and embryos and the development of transgenic fish with enhanced production characteristics. Genetic and endocrine technologies for the production of monosex male and female fish stocks are also described.

The closing chapter summarizes the discussion of each topic at the workshop, provides recommendations to industry and describes priorities of research and development. Researchers as well as teaching faculty in the aquaculture field will find this volume of great value.

Dec 2, 2012

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Reproductive Biotechnology in Finfish Aquaculture - Elsevier Science

Reproductive Biotechnology in Finfish Aquaculture

Proceedings of a Workshop hosted by the Oceanic Institute Hawaii, USA, in Honolulu, 4–7th October 1999

First Edition

Cheng-Sheng Lee

The Oceanic Institute, Waimanalo, Hawaii 96795, USA

Edward M. Donaldson

Department of Fisheries and Oceans, West Vancouver Laboratory, West Vancouver B.C., CANADA V7V 1N6



Amsterdam – London – New York – Oxford – Paris – Shannon – Tokyo

Table of Contents

Cover image

Title page

Copyright page

Reproductive Biotechnology in Finfish Aquaculture

Preface and acknowledgements

Effects of stress on fish reproduction, gamete quality, and progeny


1 Introduction

2 Effects of stress on adult physiology and quality

3 Variation in the physiological and organismic stress response

4 A progeny-protecting system: a modest proposal

5 Concluding remarks

Effect of broodstock nutrition on reproductive performance of fish


1 Introduction

2 Effect of food restriction

3 Effects of nutrition on fecundity of broodstock fish

4 Effect of broodstock nutrition on fertilization

5 Effect of broodstock nutrition on embryo development

6 Effects of broodstock nutrition on larval quality

7 Timing of broodstock nutrition

8 Valuable ingredients for broodstock diets

9 Broodstock feeding practices

10 Concluding remarks


Genetics and broodstock management of coho salmon


1 Introduction

2 Materials and methods

3 Results

4 Discussion

The environmental regulation of maturation in farmed finfish with special reference to the role of photoperiod and melatonin

1 Introduction

2 Seasonally-changing light regimes

3 Constant photoperiod regimes

4 Direction of change of daylength and photoperiodic history

5 Endogenous biological clocks or rhythms

6 Photoperiod–temperature interactions

7 Effects of salinity

8 Effects of fish size/nutritional status

9 Melatonin secretion and the timing of reproduction

10 Transduction of photoperiodic information to the reproductive axis

11 Application of photoperiod techniques to aquaculture

12 Overall conclusions


Endocrine manipulations of spawning in cultured fish: from hormones to genes


1 Introduction

2 Reproductive dysfunctions of cultured fish

3 Hormonal induction of ovulation and spawning

4 Next generation of spawning induction therapies

5 Conclusions


Disease prevention and control for gametes and embryos of fish and marine shrimp


1 Introduction

2 Vertically transmitted pathogens

3 Methods for prevention and control

4 Saprophytic infections of embryos during incubation

5 Cryopreservation of gametes

6 Summary and conclusions


Cryopreservation of finfish and shellfish gametes and embryos


1 Introduction

2 Cryopreservation research—Taiwan as an example

3 Recent advances in cryopreservation research

4 Cryopreservation research directions

5 Epilogue


The role of aquatic biotechnology in aquaculture


1 Introduction

2 Discussion

3 Conclusion


Genetic improvement of aquaculture finfish species by chromosome manipulation techniques in Japan


1 Introduction

2 Application of polyploid techniques for aquaculture

3 Application of gynogenetic and androgenetic techniques to aquaculture

4 The use of chromosome manipulations to investigate sex-determining systems

5 Regulation of the use of chromosomally manipulated fish in Japan

6 Final remarks for further integration with molecular techniques


Endocrine sex control strategies for the feminization of teleost fish


1 Introduction

2 Sex in fish

3 Feminization strategies

4 Hormonal treatments

5 Clearance of steroids

6 Evaluation of oestrogen effects

7 Selected families

8 Regulatory issues

9 Recommendations for treating new species

10 Conclusions and future prospects


Monosex male production in finfish as exemplified by tilapia: applications, problems, and prospects


1 Introduction

2 Natural variation in sex ratio

3 Benefits of monosex culture

4 Methods for production of monosex males

5 Production of genetically male Nile tilapia monosex (GMT)

6 Monosex male culture in other species

7 Conclusions: problems and potential for the future


General discussion on Reproductive biotechnology in finfish aquaculture

1 Introduction

2 Stress

3 Broodstock nutrition

4 Genetics

5 Environment

6 Endocrine regulation of spawning

7 Disease prevention and control

8 Cryopreservation

9 Biotechnology/transgenics

10 Chromosome manipulation

11 Sex control

Keyword Index



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Reproductive Biotechnology in Finfish Aquaculture

Proceedings of a Workshop hosted by the Oceanic, Institute Hawaii, USA, in Honolulu, 4–7th October 1999



The Oceanic Institute, 41-202 Kalanianaole Hwy., Waimanalo, Hawaii 96795, USA


Department of Fisheries and Oceans, West Vancouver Laboratory, 4160 Marine Drive, West Vancouver B.C., CANADA V7V 1N6

Preface and acknowledgements

Techniques for controlling and manipulating the reproduction of aquatic species to increase aquaculture production have been the subject of a number of studies throughout the world. The Aquaculture Interchange Program, funded by the National Oceanic and Atmospheric Administration (NOAA), Grant #NA86RG0041, at the Oceanic Institute, Hawaii (OI) conducted a workshop on Reproductive Biotechnology in Finfish Aquaculture in October 1999 in Honolulu, Hawaii. The objectives of the workshop were to review progress and assess the status of biotechnology research that can be applied to control the reproduction of species for aquaculture, to facilitate the exchange of information among researchers, to elucidate future research needs for controlling reproduction, and to make recommendations to the industry for ways to safely use reproductive biotechnology to the best advantage.

This special issue is the proceedings of this workshop. The papers that were presented included both original research and reviews. Information on secure sources of desirable fertilized gametes and embryos, techniques for manipulating maturation and spawning, and methods of preventing the transmission of diseases were presented. Factors that influence reproductive performance, such as stress, broodstock nutrition, and the environment were also considered. Species discussed at the workshop included Coho salmon, rainbow trout, turbot, Japanese flounder, seabream, striped bass, eel, tilapia, and loach. The research reported was carried out in Canada, Japan, Singapore, Spain, Taiwan, the UK, the USA, and the Philippines.

This workshop was planned and organized by Cheng-Sheng Lee (OI) and Edward Donaldson. Financial support to cover the expenses of holding the workshop in Honolulu and for the costs of publishing the proceedings was generously provided by NOAA. The organizers would like to thank Niall Bromage for his help in the final editing of this special issue. They would also like to express their appreciation to the invited speakers for their participation in the workshop and for the time they invested in writing the papers. The content of these papers reflect the opinions of the authors and not necessarily those of NOAA. The organizers would also like to thank Pat O’Bryen for her assistance in editing and preparing the manuscripts for publication.

Cheng-Sheng Lee, The Oceanic Institute, 41-202 Kalanianaole Hwy., Waimanalo, HI 96795, USA

Effects of stress on fish reproduction, gamete quality, and progeny

Carl B. Schreck*; Wilfrido Contreras-Sanchez; Martin S. Fitzpatrick    Oregon Cooperative Fish and Wildlife Research Unit,¹ U.S.G.S. and Department of Fisheries and Wildlife, 104 Nash Hall, Oregon State University, Corvallis, OR 97331-3803, USA

* Corresponding author.

¹ Supported cooperatively by Oregon State University, the Oregon Department of Fish and Wildlife, and the U.S.G.S.


Different taxa of fish have different tolerances to stress. This implies that for a particular stressor, severity may vary depending on the species to which it was applied. Species may differ in the nature of their physiological response and reproductive consequences to stressors. For example, disturbance or handling may affect the timing of reproduction—accelerating or delaying it as the case may be—in species such as rainbow trout (Oncorhynchus mykiss); however, tilapia (Oreochromis niloticus) respond by acceleration or complete inhibition of reproduction, depending on the maturational stage when the stressor is experienced. Strategies for coping with stress affect reproductive fitness either in terms of gamete or progeny quality. The physiology associated with maturation and spawning appears tightly coupled with stress physiology. Environmental variables, particularly nutrition, are ultimately important in affecting gamete quality and reproductive timing. The physiological response to stressors is also quite polymorphic, within and between species. For example, the circulating concentration of the primary stress response factor cortisol varies greatly among resting and among stressed rainbow trout stocks. Immunocapacity can be influenced by stress, reducing reproductive fitness of broodfish. We propose that maternal systems have been developed to buffer eggs from deleterious consequences of stressors, including regulation of transfer of substances of maternal origin to the egg and in mechanisms controlling the timing of reproduction. Effects of nutritional stressors are moderated by effects on timing of first maturity or subsequent reproductive events and/or by maintenance of quality of some eggs via atresia of others. Deleterious overload of eggs with substances such as cortisol is likely prevented by limiting entry of these compounds into the eggs. Barriers to vertical transmission of numerous pathogens seem to exist, while maternally derived immune protection is provided to assist with disease prevention of pathogenic organisms acquired from parents or by direct post-spawning infection. Timing of reproductive events including puberty, atresia, maturation and ovulation are influenced by other physiological variables responsive to stressors. Knowledge of how a stressor might affect the physiology of a species can help in development of management tactics that lessen the impact of a stressor or even in the development of therapeutants. Published by Elsevier Science B.V.



Fish reproduction

Gamete quality


Received 1 October 2000; accepted 31 December 2000

1 Introduction

Adult fish are exposed to stressful situations in the wild as well as under culture conditions. The nature of the physiological responses to threats seems stereotypic at a gross level amongst life history stages once fish have begun feeding exogenously. However, the stress response can be polymorphic with regard to species of fish, stage of maturity, and type and severity of stressor (for general discussion of these topics in fish, see Donaldson, 1981; Schreck, 1981, 2000; Barton and Iwama, 1991; Wendelaar-Bonga, 1997; and for discussion of stress and reproduction, see Pankhurst and Van Der Kraak, 1997). The word stress has been defined in several ways (Pickering, 1981). Here, a physiological nomenclature is used referring to stress as the response of the body, i.e., a physiological cascade of events that occurs when the organism is attempting to resist death or reestablish homeostatic norms in the face of an insult. It is our aim in this paper to describe the stress response of adult fish and how it relates to reproductive fitness.

Comprehending the population-level consequences of stressful events requires understanding the effects of stress on an individual; such understanding is critical for conservation biology, stewardship of wild populations, and aquaculture. From an ecological as well as a management perspective, factors affecting broodfish quality can be reflected in the number and quality of their progeny. Stressful situations can overwhelm the homeostatic mechanisms of a fish, thereby placing a load on the body in an attempt to compensate and achieve another level of stasis. When stressed, fish can assume a different mode of operating, allostasis (see Sterling and Eyer (1988) and McEwen (1998) for discussions of the concept), that is adaptive in terms of keeping the animal alive in the face of the stressor but can be maladaptive in terms of performing other necessary life functions or of reproductive fitness (Schreck, 2000). This paper briefly describes the basic physiological response to stressors and consequences on fitness of broodfish. We then review effects of stressors applied at different times during reproductive maturation and speculate about effects of stressor severity. The polymorphic nature of the stress response is exemplified by comparison of the stress-invoked concentrations of one of the primary stress factors, cortisol, among species and among stocks of a rather well-studied organism, the rainbow trout, Oncorhynchus mykiss. The immune system is then discussed in some depth since there is little or no information concerning other physiological functions of this steroid. It is not surprising that the response of broodfish to stressors in terms of physiology or reproductive tactics is extremely varied given the great diversity in reproductive modes found amongst fishes, with the extant forms divisible into 32 reproductive guilds (Balon, 1975). To help understand effects of life history phenotypes, the responses by fish with very different life histories are compared. In conclusion, a model that describes the interaction of maternal systems with eggs to provide protection of eggs from maternal stress-induced responses is proposed.

2 Effects of stress on adult physiology and quality

2.1 The physiological stress response

Most stressful situations induce a rather predictable stress response in fish similar to the General Adaptation Syndrome (Selye, 1950, 1973). The stress response begins as changes in tissue and organ function that attempt to cope with or compensate for the stressor, resulting in a move away from homeostasis—these changes are the physiological response that may differ between individuals in rate or magnitude, but share general characteristics in their mode and action. Since the elements of the physiological stress response have been described elsewhere (Donaldson, 1981; Schreck 1981, in press; Barton and Iwama, 1991; Wendelaar-Bonga, 1997), only a brief overview will be presented here, recognizing that much of our understanding of how fish respond to stressors is based on the study of juvenile life history stages. The physiological stress response happens following perception of the stressful event. Most stressors induce a neuroendocrine cascade involving an immediate release of catecholamines and activation of the hypothalamic–pituitary–interrenal axis (HPI). Complete activation of the HPI lags somewhat behind the catecholamine response because of the time needed for neural stimulation of release of the neuropeptide corticotropin releasing factor (CRF) originating from the hypothalamus that causes pituitary synthesis and secretion of corticotropic hormone (CTH), which must travel via the circulation to the interrenal to stimulate synthesis and secretion of glucocorticoid hormones (cortisol in teleosts). Together, catecholamines and glucocorticoids initiate secondary and tertiary stress response factors. The physiology of the relationships between stress and reproduction for the poikilothermic vertebrates was reviewed by Greenberg and Wingfield (1987).

One aspect of the secondary responses consists of mobilization of energy-rich substrates by depletion of hepatic glycogen stores, elevation of plasma levels of glucose, effects on circulating levels of free fatty acids, and general inhibition of protein synthesis (see Mazeaud et al., 1977; Pickering, 1981). In this regard, stress can be viewed as being anti-anabolic (Schreck, 1992). Stress is also manifested in hydromineral balance, resulting in water loading in fish in fresh water and loss in those in salt water. Electrolyte concentrations can also be affected, but it is unclear if these are direct responses to a primary stress factor or are more indirect due to water flux. Osmoregulatory consequences produce an allostatic load. Stress also encompasses various arms of the immune system; stress is generally believed to result in depressed immune capacity. There are numerous tertiary responses resultant for the primary and secondary stress responses. For example, normal behavior can be impaired (Schreck et al., 1997).

Of course, a particular stressor can also cause a specific effect or effects in fish that are the direct result of that stressor alone. Toxicants serve as good examples of stressors that have specific actions because they target specific physiological systems. In fact, certain toxicants may not activate the more general stress response system at all. Because of the highly variable effects of contaminants on the physiology of fish, this category of stressor will not be considered further in this review.

Taken together, the allostatic load due to stress appears to reduce reproductive fitness in fish (Schreck, 2000). This can be manifested directly in reduced survivorship of the adult or via reproductive failure. Reproductive impacts can be attributable to effects on maturation, time of spawning (or ovulation), gamete quality, and progeny quality. These factors will be discussed in detail subsequently.

3 Variation in the physiological and organismic stress response

Numerous factors can contribute to the variation expressed in the physiological and organismal response to stressors. These include gender, age, season, physical condition, social status, water quality, and the nature of the stressor (Schreck, 1981; Wendelaar-Bonga, 1997). The inherited performance capacity and the stress-invoked phenotype can have enormous repercussions on the way a particular fish responds to stressors (Schreck, 1981; Pottinger et al., 1992; Pottinger and Moran, 1993).

3.1 Variation in the physiological stress response

3.1.1 Primary stress response factors

As mentioned above, the physiological response to stressors can be polymorphic with considerable variation between species. Thorough integrative and comparative studies have not been undertaken to contrast how inter- and intraspecific variation are manifested in the stress response; however, there is sufficient literature to suggest that different species can respond differently to similar stressors. For example, while salmonids respond to handling and crowding stressors with an almost immediate (i.e., measurable in minutes after the onset of the stressor) elevation in circulating levels of cortisol (Donaldson, 1981; Barton and Iwama, 1991), measurable increases in cortisol levels of the sea raven, Hemitripterus americanus, are apparent only an hour after the onset of the stressor (Vijayan and Moon, 1994).

Although fish, like other vertebrates, respond to stressors by changing physiological function to reallocate energy for the purposes of coping, the actual control and nature of the physiological response shows broad phylogenetic differences. For example, while cortisol is the predominant stress steroid of teleosts, our laboratory has been unable to document the presence of cortisol or any other glucocorticoid in adult Pacific lamprey, Lampetra tridentata, even following severe handling and prolonged crowding that resulted in elevation of glucose; also, no glucocorticoid was detected in this species when injected with the radio-labeled corticosteroid precursor pregnenolone (Close, unpublished data). Variation among strains of the same species is also evident; e.g., the cortisol stress response has been shown to vary among stocks of rainbow trout (Pottinger et al., 1992; Fevolden et al., 1993; Pottinger and Moran, 1993; Pottinger and Carrick, 1999). Individual heritability for cortisol’s elevation in response to stress is moderate to high (Pottinger and Carrick, 1999).

To understand the magnitude of the physiological variation in the physiological stress response extant within a species of adults, information on cortisol concentrations in the circulation from research on rainbow trout has been compiled (Table 1). This species and this physiological variable were selected because they were the most represented in the literature. Mean resting levels of cortisol varied from < 2.0 to 540 ng/ml, and the values after stress ranged from 8 to 735 ng/ml. Some authors have detected significant differences between males and females of the same age and strain (Campbell et al., 1994; Clements, 1996; Pottinger and Carrick, 1999). In this laboratory, the overall pattern of the cortisol response to acute stressors was found to change dramatically during the period when females were undergoing vitellogenesis (Contreras-Sanchez, 1995). These trout not only had higher than expected resting cortisol levels (mean = 51.5 ng/ml), but also exhibited a very rapid, but short duration elevation in plasma cortisol levels, peaking in 20 min and returning to pre-stressor concentrations within another 20 min. Such a rapid, attenuated response may be adaptive for gamete quality (see below). Of course, apparent differences could also arise from inconsistencies among methods.

Table 1

Resting and stressor-induced circulating cortisol levels in female, male and mixed sex populations of adult (post-puberty) rainbow trout of various stocks at various times during reproductive maturation

a Quantified using a fluorimetric assay.

Steroidogenesis was suppressed when ovarian follicles of rainbow trout were exposed in vitro to physiological levels of cortisol, suggesting a possible mechanism by which stress may compromise vitellogenic processes (Carragher and Sumpter, 1990). However, these results could not be reproduced by Pankhurst et al. (1995) and Pankhurst (1998). Wendelaar-Bonga (1997) presented an excellent review of related results, pointing out the difficulty in drawing conclusions because of inconsistencies in the data; negative effects of stress on reproduction mediated by cortisol were reviewed by Greenberg and Wingfield (1987).

The physiological response of a female to stress could have considerable consequence in terms of gamete quality and progeny fitness. Experiments in guinea pigs and mice indicate that the stress response in the mother, such as elevated levels of catecholamines and corticosteroids, can be reflected in the fetus and may cause alterations to the developing offspring (Dauprat et al., 1990; Takahashi et al., 1998). Several studies have shown the presence of large reservoirs of maternally contributed hormones in mature teleostean oocytes, supporting the idea that hormones could play a major role in regulating developmental processes post-fertilization (Lam, 1985; Brown et al., 1989; Brown and Bern, 1989; Feist et al., 1990; Schreck et al., 1991; Ayson and Lam, 1993; Yeoh et al., 1996a,b). Maternal transfer of hormones such as cortisol could be modified by physiological processes induced by stress, suggesting that concentrations in eggs could be affected (Campbell et al., 1992, 1994; Yeoh, 1993). Cortisol concentrations in coho salmon, O. kisutch, eggs were significantly higher when the fish were stressed for 2 weeks prior to spawning (Stratholt et al., 1997). McCormick (1998) also found that cortisol administered to ambon damselfish, Pomacentrus ambionensis, resulted in elevated ovarian cortisol concentrations, similar to those of fish experiencing stress in the natural environment. However, we were unable to find elevation in cortisol concentrations in rainbow trout eggs that were stressed up to 3 months prior to spawning. In fact, elevated levels of maternal cortisol at ovulation were not reflected in the concentrations in ovarian fluid or eggs (Contreras-Sanchez, 1995). Furthermore, the early developing embryo has the biochemical machinery to metabolize steroid hormones, including cortisol (Yeoh et al., 1996a,b), which suggests that regulation of maternally contributed factors occurs post-fertilization in the embryo. Implantation of cortisol into adult tilapia, Oreochromis mossambicus, resulted in reduced oocytes size and circulating testosterone and 17β-estradiol concentrations (Foo and Lam, 1993).

3.1.2 Immunosuppression and disease resistance

Unless killed outright by a stressful situation, fish tend to die due to secondary rather than primary effects of the stressor in situations where they cannot compensate adequately. One such secondary effect occurs when stressed fish succumb to pathogens that they could otherwise resist. Stress is known to be immunosuppressive in fish (see review by Schreck, 1996). For adult fish, an extreme example of this is to be found with Pacific salmon, Oncorhynchus spp., that all die after spawning. Maturation and spawning in Pacific salmon is accompanied by loss of pituitary control of the interrenal secretion of cortisol; concomitant with this is a decrease in the ability to clear cortisol from the circulation (Robertson and Wexler, 1957; Hane and Robertson, 1959; Robertson et al., 1961; Hane et al., 1966; Idler et al., 1963, 1966; Donaldson and Fagerlund, 1968, 1972; Dickhoff, 1989; Stein-Behrens and Saplosky, 1992). In senescent mammals cortisol down-regulates its own receptor centrally, thereby rendering negative feedback less sensitive Stein-Behrens and Saplosky (1992). The net effect is that plasma levels of cortisol become elevated and the fish succumb to Cushing’s syndrome. Consequently, the fish either become infected with new pathogens or can no longer resist latent pathogens. Sustained elevated levels of cortisol seem to be the cause of the immunosuppression that renders the fish vulnerable to the pathogens. Any species of broodfish exposed to any stressor that elevates circulating concentrations of cortisol for more than a brief period of time could exhibit similar symptoms.

Cortisol (and stress) depresses the ability of fish leukocytes to form antibodies (Maule et al., 1987, 1989). This suppression seems to operate through a cortisol receptor-mediated system in the lymphocyte (Maule and Schreck, 1990a, 1991). Stress and cortisol also affect the numbers of leukocytes in immune organs (Maule and Schreck, 1990b). Adult fish of either sex may produce significant amounts of androgens such as testosterone during maturation that are immunosuppressive (Slater, 1991; Slater et al., 1995a) and appear to operate through a receptor-mediated system (Slater et al., 1995b). While cortisol may interfere with production of an essential cytokine (Tripp et al., 1987), androgens may be immunosuppressive by causing premature leukocyte death (Slater and Schreck, 1997). It should be noted that these conclusions about stress and the endocrine regulation of the immune system are based on studies employing juvenile tissue; work of this nature with adults is extremely limited.

Nevertheless, some research has linked cortisol to immune function in adults. In adult female chinook salmon, O. tshawytscha, numbers of antibody producing cells in peripheral blood were negatively correlated with cortisol and positively correlated to androgens and oestrogen; whereas no correlation was observed with progestin (Maule et al., 1996). In that study, fish migrating upstream and those held under hatchery conditions at warmer, variable temperatures had elevated cortisol levels and a reduced capacity to elaborate antibody-producing leukocytes. When those fish were moved to cooler, constant temperature aquaculture conditions about a month before ovulation, cortisol levels decreased and antibody producing cell numbers increased and remained high until spawning. Glucocorticoid receptors in the leukocytes did not appear to vary over the period of maturation. Lysozyme activity, which seems responsive to stressors, is similar between primary and secondary circulations in early vitellogenesis but becomes lower in the blood in chinook salmon nearer to ovulation that had returned to a hatchery (Maule et al., 1996). Extending the conclusions from the above studies, late maturation-stage broodfish may be particularly vulnerable to stress-induced disease since resting testosterone and cortisol titres would be elevated, which could affect both specific and non-specific arms of the immune system.

3.2 Temporal scaling, gamete quality, and reproductive fitness

To be able to understand the effects of stressors on a particular broodstock, one must consider the time during gonadal development at which stress occurs and the severity and duration of the stressor. These factors may lead to completely different responses depending on the species’ reproductive strategies and costs. Under stressful conditions, trade-offs between reproductive efforts, somatic growth and survival may occur. These trade-offs imply that under adverse conditions, a female can select between energy allocated for maintenance and somatic growth, or energy for reproduction. The most common trade-offs involve fecundity and life span. The considerable loss of body size or growth consequent to stressful conditions has been well documented for many species. Roff (1982) proposed two patterns of energy allocation during reproductive trade-offs: a) maintenance of body weight and adjustment in gamete production, and b) maintenance of constant numbers of eggs at the expense of somatic tissue. The former trade-off may result when stressors compromise the development of the ovary during vitellogenesis, which results in egg atresia and subsequent reabsorption. The second could happen when somatic tissue is severely affected during pre-spawning migration but number and quality of eggs remain constant. Egg number appears to evolve around selection for egg size (Fleming and Gross, 1990).

It is well established that environmental variables can affect timing of reproductive functions. The transition from juvenile to adult stage, puberty, can be difficult to describe; puberty in male fish was recently reviewed by Schulz and Goos, 1999). There is considerable plasticity for age and size of fish at sexual maturity in response to stress (Stearns and Crandall, 1984). Ultimate reproductive timing factors, particularly nutrition, can determine age to first maturity and also age at which and frequency of subsequent reproductive events in fish (Sadleir, 1973). For example, dietary protein was shown to be important for puberty and gamete quality in Nile tilapia, Ore. niloticus (Gunasekera et al., 1995). Nutritional stress experienced earlier in the life cycle can affect the timing of puberty, maturation as well as fecundity. With older-lived species, age at first spawning can be delayed by years by nutritional stress; similarly, the duration between spawning events can be prolonged by years due to nutritional deficits.

The mechanism by which nutritional state is translated into the physiology that establishes the organism’s reproductive fate remains ill defined. How does the fish establish that it is safe to proceed with further reproductive development?

Environmental stressors and particularly nutrition can also affect realized fecundity and gamete quality. In general, larger females produce larger eggs, which give rise to progeny that are believed to have an ecological advantage over those from smaller eggs. Therefore, a stressor that affects growth may lead to the production of progeny that are already at a disadvantage because of their smaller size. The number of ripe eggs a female produces is also based on environmental quality. A female produces oogonia during early developmental stages and then during the process of oogenesis establishes the actual number of these that will start to mature and that will be ovulated, with the remainder being reabsorbed through atresia. Fish must balance the production of eggs against the requirements to maintain at least some minimal level of quality (size and content). How fish weigh the trade-off between these variables is unknown. Literature in this area includes discussions on reproductive tactics relative to egg size, fecundity and age at maturity (Hislop, 1984); resource allocation for somatic growth and gonad nutrient content (Iles, 1984; Encina and Granado-Lorencio, 1997); age-structuring, energy acquisition and fitness (Ware, 1984); the relation of hatching success to female condition (Laine and Rajasilta, 1999); and the importance of nutrients, particularly thiamine, in early embryo mortality syndrome (Hornung et al., 1998).

The most fundamental decision by a fish in the face of stress is whether or not to reproduce. If reproduction is to occur, then the fish must balance fecundity with egg quality. To clarify tactics adopted by fish with very different reproductive life histories, we compared Nile tilapia (multiple spawning—throughout the year) with rainbow trout (simple spawning—once per year at most (Table 2). The period of maturation when fish are exposed to a stressor appears important. Tilapia encountering stressors (disturbance and agitation) during early ovarian development delayed ovulation while those stressed during late vitellogenesis spawned immediately (Contreras-Sanchez, unpublished data). Similarly, ovulation occurred earlier when rainbow trout were exposed to a mild stressor during late vitellogenesis (Contreras-Sanchez et al., 1998). Interestingly, ovulation was also advanced if fish were mildly stressed during the whole vitellogenic period. The severity of the stressor also appears important. Trout under more severe stress (emersions) for 9 months prior to spawning produced small eggs and delayed ovulation (Campbell et al., 1992). Under a milder stress regime (disturbance once per day) during early vitellogenesis, rainbow trout also produced smaller eggs that varied in size, while there was no effect on mean egg size in fish stressed during late vitellogenesis (Contreras-Sanchez et al., 1998). The fish mildly stressed during the whole vitellogenic period showed no effect on mean egg size but the egg sizes were more heterogeneous. Only slightly more than one-half of the oocytes of female striped trumpeter, Latris lineata, that experienced frequent handling developed past the cortical alveoli/early vitellogenic stage; the mean volume of eggs produced each day was greater in stressed than non-stressed fish (Morehead et al., 2000).

Table 2

The effects of stress at various times on reproductive traits of two species with differing reproductive strategies

Other teleosts have also been shown to employ quite varied reproductive strategies to cope with stress. For example, stress or the lack thereof seems to be involved in regulating sex reversal in hermaphroditic species where density, growth and social status are important regulators of phenotypic sex (Robertson, 1972; Shapiro, 1981; Ross, 1987; Hourigan, 1986). For live-bearers, there seems to be only anecdotal information indicating that parturition is accelerated by stress. Fish of two genera of poeciliid, Poecilia and Phallichtys, were observed on numerous occasions to release their young while unstressed individuals at similar stages of pregnancy did not (Contreras-Sanchez, unpublished data). Shifts in reproductive tactics under stressful situations are probably important for optimizing reproductive fitness for fish in the wild, and understanding of such processes is obviously important for management of wild and hatchery stocks.

3.3 Stress and progeny quality

Unfortunately, other than nutritive effects mentioned earlier, only limited information exists on how stress experienced by broodfish affects their progeny. Campbell et al. (1992, 1994) found that relatively severe stress in rainbow trout for prolonged times can affect progeny survival. This reduced progeny viability could be due to limited energetic reserves allocated to the eggs as well as mechanical damage caused by the specific stressor. In contrast, progeny from females that experienced mild stress during various stages of vitellogenesis, by contrast, did not suffer abnormal mortality (Contreras-Sanchez et al., 1998). However, one can surmise that smaller eggs and hence smaller hatchlings resulting from some stressful situations, as found by Contreras-Sanchez et al. (1998), could be maladaptive in the wild. An interesting study by McCormick (1998) suggested that stress induced by the presence of predators and to a lesser extent conspecifics in female ambon damselfish resulted in eggs with higher cortisol concentrations and that there was an inverse correlation between levels of this hormone in eggs and egg size. The implications of this work are that behavioural interactions and stress incurred by broodfish can affect progeny quality.

4 A progeny-protecting system: a modest proposal

We suggest that the females have a mechanism that allows them to protect or buffer their eggs from the deleterious effects of stress. At a gross level, such buffering can be exemplified in the trade-off between fecundity and egg quality when a female reabsorbs some eggs to spare energy or nutrients for the remainder during nutritionally stressing times. However, there are likely numerous maternal mechanisms that also spare individual eggs from potentially maladaptive physiological responses induced by stressors. Based on the extant evidence, two main avenues where the stressed maternal system can maintain the quality of eggs are proposed: (1) the quantity and nature of substances transferred from the female to the egg, and (2) the timing of maturation and ovulation (Fig. 1).

Fig. 1 Conceptualization of a buffering system that can maintain egg quality during resting and stressed states. Short up and down arrows indicate the effects of buffering on maintaining levels of some factor or process above some minimum or below some maximum (homeostasis or allostasis). The vascular system provides numerous factors to the developing egg and also to the physiological systems controlling timing of reproductive events (Reproductive Clock). For example, an overload of cortisol (F) in the egg is prevented by corticosteroid binding protein (CBP), brevity of the stress response in mature fish affecting the clock, or via enzymatic buffering converting F to cortisone (E). Nutrients, (e.g., vitellogenin, Vg) and specific antibodies (Ab) and non-specific immune factors help protect against pathogens. The clock can spare energy and nutrients for eggs via timing of ovarian atresia or by optimizing spawning timing. Central (neural) control of the Reproductive Clock is via direct or indirect (e.g., endocrine) mechanisms.

Maternal transfer of vitellogenin, lipid and other nutrients are essential during egg maturation. The composition and accumulation of yolk in teleost eggs were discussed in detail in the excellent review by Wiegand (1996). However, while certain levels of compounds such as hormones could be important for regulating development (Lam, 1985; Brown et al., 1989; Brown and Bern, 1989; Feist et al., 1990; Schreck et al., 1991; Ayson and Lam, 1993; Yeoh et al., 1996a, b), an overabundance could be detrimental. For example, cortisol is secreted when fish are stressed and it is anti-developmental, anti-growth (Schreck, 1992), and immunosuppressive (Schreck, 1996). Furthermore, Cloud (1981) found that hatching was accelerated when medaka, Oryzias latipes, eggs were incubated with deoxycorticosterone. It is thus not surprising that at ovulation both stressed as well as unstressed rainbow trout had 17 times less cortisol in ovarian fluid and 30 times less in eggs than in their circulation (Contreras-Sanchez, 1995). It is thus likely that the female protects the eggs from hypercortisolism.

We postulate that this could be accomplished by three

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